Methods for thrombolytic therapy

ABSTRACT

Certain Factor XIIIa inhibitor compounds have been discovered which have been found to be useful in the lysis of blood clots and thus adaptable for administration in thrombolytic therapy either alone or together with plasminogen activator.

This is a division of application Ser. No. 07/032,123, filed Mar. 27,1987, abandoned.

BACKGROUND OF THE INVENTION

Thrombosis, an excessive formation of clot within a blood vessel, givesrise to thrombotic strokes, deep vein thrombosis, myocardial infarctionand other medical conditions which may result in necrosis of tissues andoftentimes death of a patient. Even if death does not occur, thromboticattacks are accompanied by damage to cells to which circulation has beenprevented by thrombi formation. Removal of the thrombi by lysis isessential and the rate of lysis may be critical in ultimate patientrecovery.

Lysis may occur normally in hours or days by the action of a proteolyticenzyme, plasmin, which is present in plasma as the inactive precursor,plasminogen, and which is activated by plasminogen activators, such as(pro)urokinase, urokinase or tissue plasminogen activator. Since theoccurrence of a thrombotic event calls for rapid remedial action,administration of exogenous tissue plasminogen activator or(pro)urokinase is currently looked to in thrombolytic or fibrinolytictherapy. However, a still further reduction in lysis time is necessaryto minimize cell injury.

Factor XIIIa is an enzyme responsible for the final event in thecoagulation of blood. It is a plasma or platelet transglutaminase whichis the activated form of Factor XIII, also known asfibrin-stabilizing-factor. It is essential for normal hemostasis and isresponsible for the cross-linking of fibrin. This step is sometimesdescribed as the transformation of soft clot to hard clot.

STATEMENT OF THE INVENTION

According to the present invention it has been discovered that byadministering a compound hereinafter defined which is a Factor XIIIainhibitor, blood clots may be more rapidly lysed. Further, it has beendiscovered that when administered together with tissue plasminogenactivator, the time and extent of lysis is increased multifold in vitro.Thus, the discovery has provided a method for thrombolytic orfibrinolytic therapy which comprises administering a therapeuticallyeffective amount of a Factor XIIIa inhibitor compound, alone or inadmixture with a tissue plasminogen activator or a combination ofplasminogen activators.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based on the discovery that certain compoundswhich have been found to inhibit the action of Factor XIIIa, a plasmatransglutaminase enzyme which catalyzes a number of reactionsstabilizing blood clots, have been found unexpectedly to have theproperty of making blood clots more susceptible to being lysed either atphysiological levels or administered levels of plasminogen activator. Ithas further been discovered that when a Factor XIIIa inhibitor isemployed together with a plasminogen activator, such as tissueplasminogen activator an enzyme which converts plasminogen to plasmin, amultifold increase in the rate and extent of lysis may be achieved.Thus, there is provided a method useful in fibrinolytic therapy whichcomprises administering a Factor XIIIa inhibitor to a patient sufferingfrom thrombosis or susceptible to thrombotic attack. The method may beemployed in acute therapy for immediate and rapid lysis of clots byadministering a Factor XIIIa inhibitor together with plasminogenactivator to a patient suffering from a thrombotic attack. It is alsouseful for continued therapy after initial relief from thrombotic attackhas been achieved by providing a more complete lysis of the clot andthereby minimizing complications from reocclusion. The method also maybe useful as a prophylactic means by administering a Factor XIIIainhibitor alone to a patient considered to be in a high risk thrombosiscategory.

One class of Factor XIIIa inhibitor useful in the practice of thepresent invention is an acyloxiranecarboxylic acid compound representedby the formula ##STR1## and pharmaceutically acceptable salts. In thisand subsequent formulas R is alkyl, alkenyl or aralkyl;

X is --NHR' or OR" wherein R' and R" independently are hydrogen or loweralkyl.

As employed in the specification and claims, the expressions for thegroups have significances hereinafter detailed.

By the expression "alkyl" is meant from 1 to about 10 carbon atoms. By"lower alkyl" is meant from 1 to 5 carbon atoms.

By "alkenyl" is meant an alphatic chain of 2 to about 10 carbon atomswith at least one and up to three double bonds.

By "aralkyl" is meant benzyl or phenylethyl, optionally substituted withalkyl, halo, or lower alkoxy. "Halo" embraces chloro, bromo, fluoro andiodo.

The salts contemplated are primarily sodium and potassium.

The compounds useful in the present invention must be of the cisconfiguration with respect to the epoxide ring as indicated in FormulaI.

The compounds which are amides, i.e., X is --NHR' may be represented as##STR2##

The compounds which are acids or esters, i.e., X is --OR" may berepresented as ##STR3##

The preferred acyloxiranecarboxylic acid compounds in the practice ofthe present invention are compounds which are amides, represented byFormula IA. An especially preferred compound is an antibiotic known ascerulenin having antifungal properties and represented by the formula##STR4## The compound is also known by the chemical name3-(1-oxo-4,7-nonadienyl)-oxiranecarboxamide or(2R,3S)-2,3-epoxy-4-oxo-7E,10E-dodecadienamide.

The acyloxiranecarboxylic acid compounds to be employed in the presentinvention may be crystalline solids or high boiling liquids. They aregenerally of low solubility in water, but of high solubility in solventssuch as dimethyl sulfoxide (DMSO), dimethylformamide (DMF) and mostcommon organic solvents.

Another Factor XIIIa inhibitor useful in the practice of the presentinvention is a heteroarylacrylic acid compound represented by theformula ##STR5## and its pharmaceutically acceptable salts.

In this and subsequent formulas, ##STR6## wherein Y is --NH--, --NR₂,--S--, or --O-- wherein R₂ is lower alkyl;

W is hydrogen lower alkyl, halo, or lower alkoxy; and

Z is --OR₃ or --NR₄ R₅ wherein R₃ is hydrogen or lower alkyl, and R₄ andR₅ are independently hydrogen or lower alkyl or together are --(CH₂)₅ --or --(CH₂)₂ --O--(CH₂)₂ --. By "lower alkyl" or "lower alkoxy" as aboveemployed is meant a group having from 1 to 5 carbon atoms.

The compounds which are acids or esters, i.e., Z is --OR₃ may berepresented by the formulas ##STR7##

The compounds which are amides, i.e., Z is --NR₄ R₅ may be representedby the formulas ##STR8##

The most preferred heteroarylacrylic acid compound is3-(benzimidazol-2-yl)acrylic acid represented by the formula ##STR9##

The heteroarylacrylic acid compounds are also generally crystallinesolids or high boiling liquids of low solubility in water but soluble inorganic solvents.

A compound which has been found to be especially useful is2-(1-acetonylthio)-5-methylthiazolo(2,3-b)-1,3,4-thiadiazoliumperchlorate ("thiadiazolium perchlorate compound") which may berepresented by the following formula: ##STR10##

Compounds useful in the fibrinolytic or thrombolytic therapy of thepresent invention are not limited to those of the foregoing structuralclasses. Suitable compounds may be identified initially by the FactorXIIIa inhibitor assay hereinafter described. Compounds which would besuitable in thrombolytic therapy generally exhibit at least 50 percentinhibition at a concentration of 1×10⁻⁵ M.

The plasminogen activator compounds suitable in the practice of thepresent invention include tissue plasminogen activator, prourokinase(single chain urokinase), urokinase (dual chain urokinase),streptokinase and eminase (FDC Reports, Sept. 22, 1986, T&G, page 4) asthe more important plasminogen activators. The plasminogen activatorsmay be those isolated from natural sources or produced by recombinanttechnology and include the genetically engineered variants thereof.

The acyloxiranecarboxylic acid compounds (Formula I) may be prepared bya sequence of reactions hereinafter described. Cerulenin itself may beobtained by fermentation according to Japan patent 21,638 (C.A. 73,108271k (1970)) or by any of the chemical syntheses described in theliterature. A recent synthesis by T. Ohta et al., is found inHeterocycles, 24, 1137 (1986).

The acyloxiranecarboxylic acid compounds useful in the present inventionmay be synthesized by the following sequence of reactions: ##STR11##

The epoxylactone (A) is transformed into the compounds of Formula I bytreatment with the appropriate nitrogen base (ammonia or amine) or theappropriate hydroxy compound (water or alcohol) to obtain the desiredamide, ester or acid.

When the desired compound is an amide, the epoxylactone is treated withexcess alcoholic ammonia or amine, preferably methanolic ammonia oramine. After completion of the reaction, unreacted ammonia or amine, andsolvent are removed by evaporation and the product recovered as residue.The product recovered generally is the cyclic form which may beconverted to the open form represented by Formula IA by passing theproduct through a silica gel column.

In carrying out the reaction, a methanol solution of the epoxylactone(A) is added to a solution of about 16 percent ammonia or amine inmethanol at about 0° C. After completion of the addition, the mixture isstirred at 0° C. for about 1.5 hours and then the ammonia or amine andsolvent are evaporated to obtain a residue. The latter ischromatographed on silica gel using 10 percent diethyl ether inmethylene chloride to obtain compound of Formula IA.

The compounds which are esters, i.e., X is --OR", may be prepared by thereaction of the epoxylactone (A) with an appropriate alcohol (R"OH).

In carrying out the reaction, the epoxylactone is stirred with excessalcohol R"OH for about one hour, and the excess alcohol vaporized off atreduced pressure to obtain the ester product (Formula IB-2) as residue.The latter may be purified by chromatographing on silica gel usingmethylene chloride/hexane or ethyl acetate/hexane as eluting agent.

The compounds which are acids, i.e., X is OH, may be prepared bytreating the epoxylactone with alkali to obtain an alkali salt of theacid which is thereafter carefully acidified to obtain the acid (FormulaIB-1).

The starting epoxylactone may be prepared from furfuryl alcoholemploying methods similar to that described by above cited Ohta et alfor the preparation of cerulenin, which reference is incorporated hereinby reference. Briefly, furfuryl alcohol is lithiated with an appropriatelithiating agent such as n-butyl lithium. The lithiated furfuryl alcoholis then reacted with the appropriate RI compound, i.e., an alkyl,alkenyl or aralkyl iodide to obtain 5-R-furfuryl alcohol. The ##STR12##5-R-furfuryl alcohol is then oxidized to a 6-hydroxy-6-R-2H-pyran-3-one,a hemiketal. This may be carried out conveniently by a Sharplessoxidation using for example, vanadyl acetylacetonate andtert-butylhydroperoxide. The 6-hydroxy-6-R-2H-pyran-3-one is thenoxidized to a lactol, 4-hydroxy-4-R-2-buten-4-olide. Periodic acid issuitable for this step. The lactol then may be reacted with trimethylorthoformate and tin (IV) chloride to obtain a ##STR13##methoxybutenolide which is epoxidized to the starting epoxylactone withsodium hypochlorite in dimethylformamide or a mixture of it with ether.In carrying out each of the steps, conditions similar to that detailedin the preparation of cerulenin may be employed with appropriatemodifications or substitutions as desired and within the knowledge ofthe skilled in the art.

The heteroarylacrylic acid compounds (Formula II) useful in the presentinvention are readily obtained from the corresponding knownheteroarylpropanoic acids as starting materials by oxidation ordehydrogenation, or from heteroaryl-3-hydroxypropanoic acids bydehydration ##STR14## to obtain the appropriate acrylic acid from whichesters and amides are made employing conventional procedures.

When the heteroarylacrylic acid is obtained from the heteroarylpropanoicacid by chemical oxidation, suitable oxidizing agents include mercuricacetate, mercuric oxide, potassium permanganate, manganese dioxide,copper oxide, copper sulfate, hydrogen peroxide, ferric chloride, ferricsulfate and the like. Slight excess of the oxidizing agent generally isemployed and the reaction is carried out in an organic solventappropriate for the particular oxidizing agent employed and within theknowledge of the one skilled in the art.

In a representative preparation using mercuric acetate as oxidizingagent, a solution of the appropriate heteroarylalkanoic acid in glacialacetic acid is heated with a solution of mercuric acetate in glacialacetic acid for several hours. At the end of this period, the productmay be recovered by first removing glacial acetic acid under reducedpressure, then dissolving the residue in acid, precipitating the mercurywith hydrogen sulfide, filtering off the mercuric sulfide and recoveringthe product from the filtrate by successive appropriate treatments withorganic solvent and water.

When the heteroarylacrylic acid is obtained from heteroarylpropanoicacid by dehydrogenation, the reaction may be carried out in the presenceof suitable catalysts such as nickel, copper, platinum and palladiumusing conditions appropriate for the particular catalyst. However,chemical methods of oxidation are preferred.

When the desired acrylic acid compound is to be obtained by thedehydration of heteroaryl-3-hydroxypropanoic acid, it may be carried outsimply by heating the hydroxypropanoic acid compound under reducedpressure and elevated temperatures. Temperatures in the range of150°-200° C. at 3 to 4 mm. pressure are satisfactory.

In carrying out the reaction, unmodified heteroaryl-3-hydroxypropanoicacid is heated in an appropriate vessel for such time as necessary forcompletion of the dehydration as seen by the cessation in the completionof the evolution of water vapor. The resulting acrylic acid compound maybe recovered by dissolving in dilute sodium carbonate and thenreprecipitating with hydrochloric acid.

The esters (Formulas IIA-1 and IIA-2) may be prepared from the acid bypreparing the acid chloride with thionyl chloride and then reacting theacid chloride with the appropriate R₃ OH. If a methyl ester, it may beprepared directly from methanol in the presence of hydrogen chloride.

The amides (Formulas IIB-1 and IIB-2) may be prepared from the acid bypreparing the acid chloride with thionyl chloride and then reacting theacid chloride with the appropriate amine. The reaction may be carriedout in an inert solvent such as toluene, benzene and the like.

2-(1-Acetonylthio)-5-methylthiazolo(2,3-b)-1,3,4-thiadiazoliumperchlorate is a crystalline solid which may be prepared in accordancewith the following sequence of reactions: ##STR15## The preparation andproperties of the compound is fully described in a Ph.D. thesis entitled"Thiazolo[2,3-b]thiazolium and Analogous Cations" by William JonasJones, Jr. submitted to Duke University and which is incorporated byreference.

Briefly, 1,3,4-thiadiazole-2,5-dithiol (A') is added to absolutemethanol containing sodium metal. Chloroacetone is then added and theresulting mixture stirred together to obtain the bis(acetonylthio)compound (B') as a precipitate. The latter is recovered and then heatedwith concentrated sulfuric acid at about 100° C. and thereafter themixture treated with perchloric acid and cooled to 0° C. to obtain thedesired thiadiazolium perchlorate compound (III).

The usefulness of the Factor XIIIa inhibitors for enhancing the rate ofclot lysis catalyzed by plasminogen activators may be demonstrated firstby establishing the inhibitory potencies of the compounds in a FactorXIIIa assay. Then a subsequent assay is used to determine the rate ofclot lysis.

The Factor XIIIa inhibitor assay is based on the incorporation of ¹⁴C-putrescine into casein catalyzed by Factor XIIIa. The assay is carriedout employing the procedure described in Methods in Enzymology, Vol. 45,Ch 15., pages 177-191 (1976) and using Factor XIII (F XIII) isolatedfrom human plasma. The procedure is summarized briefly and schematicallyillustrated as follows: ##STR16##

Factor XIII assay mixtures are prepared by adding stepwise, appropriateprepared solutions of thrombin and dithiothreitol (DTT) to a mixturecomprising Factor XIII in glycerol/water andtris(hydroxymethyl)aminomethane hydrochloride (Tris.HCl). To a portionof the mixture is added calcium chloride as source of calcium ionsrequired for enzyme activity and to the remaining mixture is addedinstead of calcium ions ethylenediaminetetraacetic acid (EDTA) whichserves as a blank for background.

A substrate mixture is prepared from ¹⁴ C-putrescine andN,N-dimethylcasein.

The assay tubes and control tubes are charged with the substrate mixtureand incubated at 37° C. for 20 minutes. Samples are withdrawn from eachtube, spotted onto a filter disk which is then immersed in ice coldtrichloroacetic acid solution to precipitate the casein on the filter.The filter is then washed to remove unincorporated or free ¹⁴C-putrescine and after drying is counted for ¹⁴ C-putrescineincorporated to casein from which percent activity and/or inhibition canbe calculated.

The results with cerulenin, 3-(benzimidazol-2-yl)acrylic and thethiadiazolium perchlorate compound were as follows:

    ______________________________________                                        Inhibitor          Percent                                                    Concentration      Inhibition                                                 ______________________________________                                        3-(Benzimidazol-2-yl)-                                                        acrylic acid                                                                  1.3 × 10.sup.-5 M                                                                          91                                                         4.5 × 10.sup.-6 M                                                                          78                                                         1.5 × 10.sup.-6 M                                                                          52                                                           5 × 10.sup.-7 M                                                                          19                                                         1.7 × 10.sup.-7 M                                                                           9                                                         Cerulenin                                                                     6.2 × 10.sup.-5 M                                                                          100                                                        6.2 × 10.sup.-6 M                                                                          64                                                         6.2 × 10.sup.-7 M                                                                          22                                                         Thiadiazolium perchlorate                                                     compound                                                                      1.4 × 10.sup.-5 M                                                                          100                                                          5 × 10.sup.-6 M                                                                          100                                                        1.6 × 10.sup.-6 M                                                                          99                                                           5 × 10.sup.-7 M                                                                          84                                                           2 × 10.sup.-7 M                                                                          24                                                           6 × 10.sup.-8 M                                                                           9                                                         ______________________________________                                    

The usefulness of Factor XIIIa inhibitor compounds in lysing clotsand/or inhibiting clot formation may be demonstrated in an in vitro clotlysis assay.

In the assay, whole blood or plasma is charged with ¹²⁵ I-fibrinogen.Thrombin is added to initiate clot formation and subsequently incubatedfor 2 hours to complete the cross-linking by endogenous Factor XIII.Factor XIIIa inhibitor is added prior to clot initiation or at varioustimes during the 2-hour incubation. At the end of this period, thelabelled clot is washed and lysis is initiated by exogenous tissueplasminogen activator (TPA). Exogenous plasminogen is employed in someexperiments. Endogenous plasminogen is used for others. Clot dissolutionas a function of time is monitored by the release of solubilized ¹²⁵I-fibrin-derived fragments.

In one set of experiments with no exogenous plasminogen, TPA wasemployed as the plasminogen activator at different levels with andwithout 200 μM cerulenin as the factor XIIIa inhibitor. The results seenin FIG. 1 show that there are enhanced rates and extents of lysis in thepresence of cerulenin at both physiological and pharamcological levelsof TPA.

In another set of experiments, 12 μg/mL of exogenous glu-plasminogen(glutamic plasminogen) was employed to enhance the rate and extent ofTPA catalyzed clot lysis. Under these conditions in the presence of 200μM cerulenin, lysis rates were further enhanced in a relative similarway as seen in FIG. 2.

In still another set of experiments with no exogenous plasminogen, TPAwas employed as the plasminogen activator at different levels with andwithout the thiazolium perchlorate compound as the factor XIIIainhibitor compound. The results seen in FIG. 3 show that there areenhanced rates and extents of lysis in the presence of the thiazoliumperchlorate compound at both physiological and pharmacological levels ofTPA.

In FIGS. 1 and 2, the open symbols indicate absence of cerulenin and theclosed symbols indicate the presence of 200 μM cerulenin at the time ofclot formation. Clot lysis is conducted in buffer, pH 7.5; 5 ng/mL TPArepresents approximate physiologic concentration. 50 to 5,000 mg/mLrepresents pharmacologic concentration range. FIG. 1 studies did notemploy exogenous glu-plasminogen. FIG. 2 studies employedglu-plasminogen levels at approximately 10 percent physiologic level (12μ/mL).

The process of the present invention for enhancing clot lysis and/or forinhibiting clot formation comprises intravenously administering atherapeutic dose of a Factor XIIIa inhibitor compound in a compositioncomprising the same. In general, the dose may be that sufficient toprovide between about 2 micrograms per kilogram of body weight per dayto about 100 milligrams/kilogram/day while considering patient's health,weight, age and other factors which influence drug response. The drugmay be administered either by a single injection, multiple injections orcontinuous infusion.

In the preferred process of the present invention, Factor XIIIainhibitor compound is administered with a plasminogen activator in acombination therapy. When Factor XIIIa inhibitor compound andplasminogen activator are employed in a combination therapy, it is mostdesirable to use plasminogen activator in the dose range of 5 to 40,000I.U./kg/hr and Factor XIIIa inhibitor compound in the range of 10 μg-10mg/kg/hour.

When combination therapy is employed, it is preferable to administer theFactor XIIIa inhibitor compound first in a single bolus and thereafterto administer the plasminogen activator by continuous infusion.Alternatively, it may be administered simultaneously as a continuousinfusate. Under certain circumstances it may be desirable to administersubsequent to the administration of the plasminogen activator. However,it is intended that the method of the present invention embraceconcurrent administration as well as sequential administration in anyorder.

When the Factor XIIIa inhibitor compound is employed alone, particularlyfor a prophylactic purpose, it may be employed in the range of from 10μg to 100 mg/kg/day and administered either orally or parenterally.

Compositions to be employed in the practice of the present inventioncomprises a Factor XIIIa inhibitor compound in sterile physiologicallyacceptable media such as physiological saline. Such compositions mayalso contain other ingredients for purposes such as for aidingsolubility or for preservation or the like, said ingredients beingacceptable for intravenous administration. The compositions may beprepared as concentrate compositions which may be appropriately dilutedto the appropriate treating composition immediately prior toadministration. A therapeutic composition as a unitary dose form maycontain from 2 μg to 1000 mg of Factor XIIIa inhibitor compound and asconcentrate composition may contain up to 10 grams of the compound.Compositions suitable in the preferred practice of the present inventionof co-administering plasminogen activator and Factor XIIIa inhibitorcompound may contain from 50,000 to 2 million I.U. of TPA and from 100mg to 7 grams of Factor XIIIa inhibitor compound.

The preferred compositions are those in which the Factor XIIIa iscerulenin,2-(1-acetonylthio)-5-methylthiazolo(2,3-b)-1,3,4-thiadiazoliumperchlorate or 3-(benzimidazol-2-yl)acrylic acid.

Parenteral compositions may be prepared employing the foregoingpreferred compounds or one of the following compounds as activeingredient in a manner hereinafter described. These compositions aremerely illustrative and are not to be construed as limiting.

3-(1-oxo-7-nonaenyl)oxiranecarboxamide

3-(1-oxo-nonanyl)oxiranecarboxamide

3-(1-oxo-6-phenylpropyl)oxiranecarboxamide

N-Methyl-3-(1-oxo-4,7-nonadienyl)oxiranecarboxamide

n-Propyl 3-(1-oxo-4,7-nonadienyl)oxirancecarboxylate

3-(5-methylbenzimidazol-2-yl)acrylic acid

3-(5-isopropoxybenzimidazol-2-yl)acrylic acid

3-(5-chlorobenzimidazol-2-yl)acrylic acid

3-(Imidazol-2-yl)acrylic acid

3-(Oxazol-2-yl)acrylic acid

3-(Thiazol-2-yl)acrylic acid

3-(Benzoxazol-2-yl)acrylic acid

3-(Benzthiazol-2-yl)acrylic acid

PARENTERAL COMPOSITION

One liter of a parenteral composition comprising one of the preferredcompounds or one of the foregoing compounds may be prepared from thefollowing formulation:

    ______________________________________                                                            Grams                                                     ______________________________________                                        Active Ingredient     5.0                                                     Polysorbate 80        2.0                                                     Sodium Chloride       9.0                                                     Sodium carboxymethyl cellulose                                                                      10.0                                                    Methyl paraben        1.8                                                     Propyl paraben        0.2                                                     Water, USP q.s. to 1 liter                                                    ______________________________________                                    

The parabens, sodium chloride and carboxymethylcellulose are dissolvedin one-half the total volume of water by heating to 95° C. to obtain asolution which is then filtered and autoclaved. The polysorbate isdissolved in one-third of the total volume of water, and the resultingsolution also filtered and autoclaved. Sterile active ingredient isadded to the second solution and the mixture passed through a sterilecolloid mill to obtain a suspension of active ingredient. The firstsolution is added to the suspension with stirring then U.S.P. wateradded to 1 liter. Sterile vials are filled with the suspension whilestirring.

Oral compositions also may be prepared from one of the above-namedcompounds as active ingredient in admixture with a pharmaceuticallyacceptable carrier. Suitable carriers include for liquid compositionsinclude water, glycols, oils, alcohols, flavoring agents, preservatives,coloring agents and the like; for solid preparations, starches, sugars,diluents, granulating agents, lubricants, binders, disintegrating agentsand the like. Because of their ease in administration, tablets andcapsules represent the most advantageous oral dosage unit form, in whichcase solid pharmaceutical carriers are obviously employed.

A representative composition is the following:

ORAL COMPOSITION

5000 compressed tablets, each containing as active ingredient 100milligrams of active ingredient are prepared from the followingformulation:

    ______________________________________                                                            Grams                                                     ______________________________________                                        Active Ingredient     500                                                     Starch                700                                                     Dibasic calcium phosphate hydrous                                                                   5000                                                    Calcium stearate       25                                                     ______________________________________                                    

The ingredients are finely powdered, mixed well, and then granulatedwith 10 percent starch paste. The granulation is dried and compressedinto tablets using starch as a disintegrant and calcium stearate aslubricant.

What is claimed is:
 1. A method for lysing clots in thrombotic patientswhich comprises intravenously administering a therapeutically effectiveamount of a composition comprising a Factor XIIIa inhibitor compound,said Factor XIIIa inhibitor compound being a heteroarylacrylic acidcompound represented by the formula ##STR17## and its pharmaceuticallyacceptable salts wherein Y is --NH, --NR₂, --S--, --O--, where R₂ islower alkyl;W is hydrogen, lower alkyl, halo or lower alkoxy; and Z is--OR₃ or --NR₄ R₅ wherein R₃ is hydrogen or lower alkyl and R₄ and R₅are independently hydrogen or lower alkyl.
 2. A method according toclaim 1 wherein in the heteroarylacrylic acid Factor XIIIa inhibitorcompound of the formula ##STR18## Y is NH or NR₂.
 3. A method accordingto claim 2 wherein in the formula of the heteroarylacrylic acidcompound, Z is OR₃ wherein R is H or lower alkyl.
 4. A method accordingto claim 3 wherein R is H.
 5. A method according to claim 4 wherein thecompound is 3-(benzimidazol-2-yl)acrylic acid.
 6. A method according toclaim 1 wherein plasminogen activator is also administered.
 7. A methodaccording to claim 6 wherein the plasminogen activator is selected fromthe group consisting of tissue plasminogen activator, urokinase,prourokinase, streptokinase and eminase.
 8. A method according to claim7 wherein the plasminogen activator is tissue plasminogen activator. 9.A method according to claim 1 wherein from 2 μg/kg/day to 100 mg/kg/dayof heteroarylacrylic acid compound is employed.
 10. A method accordingto claim 9 wherein from 5 to 40,000 I.U./kg/hr of plasminogen activatoris also administered.
 11. A method according to claim 1 wherein thetherapeutic dose is delivered by a single, multiple or continuousintravenous administration.
 12. A method according to claim 11 whereinthe therapeutic dose is delivered by continuous infusion.
 13. A methodaccording to claim 12 wherein the Factor XIIIa inhibitor compound isadministered first in a single bolus and the plasminogen activator thenadministered by continuous infusion.
 14. A method for lysing clots inthrombotic patients according to claim 11 which comprisesco-administering intravenously from 10 μg/kg/hr to 10 mg/kg/hr ofheteroarylacrylic acid compound and 5 to 40,000 I.U./kg/hr of tissueplasminogen activator.